Bis quaternary oximes



United rates harem G i 1 Claim. (Cl. 260-296) (Granted under Title 35, US. Code (1952), see. 266) This invention described herein may be manufactured and used by or for the Government of the United States of America for governmental purposes without the payment to use of any royalty thereon.

This is a division of application Serial No. 809,578, filed April 28, 1959.

This invention is directed to certain diquaternary pyridinium halide oximes which are useful as chemotherapeutic and pyrophylactic agents for mammals poisoned by anticholinesterases, particularly the nerve gas known as GB or sarin, i.e., isopropyl methylphosphonofluoridate.

The invention relates to 1,1-polymethylene bis (4-formylpyridinium) halide dioximes wherein the polymethylen'e group contains from 2 to 6 carbon atoms. These compounds have the structural formula E N+ on on 3 X 2- 2 J. 2

The invention also relates to 1,1-(2-butenylene) bis (4-formylpyridiniurn) halide dioximes of the formula OH=NOH OH=NOH (III) The invention further relates to 1,1-polymethylene bis (3-formylpyridinium) halide dioximes of the formula wherein R is a polymethylene group containing from 2 to 6 carbon atoms.

Another class of compounds included are the 1,1-(p- Patented Sept. 18, 1962 ice phenylenedimethylene) bis (3-formylpyridinium) halide dioximes of the formula OH=NOH This invention further relates to unsymmetrical bisquaternary 4-formylpyridinium halide monoximes of the formula CH=NOH (V) wherein R represents either three lower alkyl groups or the hydrocarbon portion of the pyridine ring.

This invention also relates to the 1,1 trimethylene bis (3-amidooximopyridinium) halides N OH 2)a 2X I C=NOH C|J=NOH NH: NH; This invention also relates to the compounds (VII) The invention also relates to the compounds While all these compounds are useful in varying degree for the purposes set out above, the different groups exhibit quite striking diiferences in effectiveness. In all the above formulas X is chloride, bromide or iodide, which appear to be equivalents as to physiological action, except for v their effect on solubility.

iodide or Z-PAM. This compound is outstanding in its ability to reactivate, in vitro, acetyl cholinesterase which has been inhibited by, for example, GB. Thus, even as compared to the very closely related 4-PAM, disclosed in Example II of the Wilson patent, 2-PAM shows much greater activity. With isopropyl methylphosphonylated acetylcholinesterase the rate constant at pH 7.4 and 25 C. in the presence of 10- M acetylcholine is 2 10 per mole per minute for 2-PAM and 1.4)(10 per mole per minute for 4-PAM. The Wilson patent shows a high rate of survival in mice which had been'poisoned with paraoxon and then treated with 2-PAM.

Nerve gas poisoning has been treated symptomatically with drugs which are pharmacologically antagonistic to esterase and also in enhancing the activity of atropine in both therapy and propylaxis. When X is bromide, the variation of the rate constant for the in vitro reactivation of GB inhibited eel acetylcholinesterase at pH 7.4 and 25 C. was found to be as follows:

Table 1 R Rate constant (llmole/minutes) (6132) 7x10 (OH2)= 6X10 (0119i 6X10 (CH2) K 1 X10 (011 5 When administered in combination with atropine to animals poisoned with GB the order of effectiveness was somewhat different. Under these conditions the compound in which R=(CH i.e., l,1'-trimethylene bis (4-formylpyridinium) bromide dioxime also known as TMB-4, was most efiective. In rats challenged with a 2 LD dose of GB administered intravenously, all of a group of six animals survived if the atropine-TMB-4 combination was administered intravenously immediately after poisoning. The atropine-Z-PAM combination saved only two of the group of animals. On the other hand, with dogs which were given a 20 LD dose of GB subcutaneously the survival ratios were the same (4/5) for the two treatments, which were given intravenously when symptoms appeared. However, the recovery time was much shorter for the surviving animals which received the TMB4, i.e., 2 hours, as against 24 hours for those receiving the 2-PAM.

A summary of the reactivation rates and survival ratios for these compounds when administered therapeutically These compounds constitute our presently preferred oup. Our compounds may be employed prophylactically, i.e., injected before exposure to the anticholinesterase agent, e.-g., GB, or therapeutically, i.e., injected subsequent to exposure.

The following series of experiments compares the effectiveness of our presently preferred compound, TMB-4, with Z-PAM applied to various animals by these two methods. The animals were poisoned by injections of GB as follows.

Mice 0:173 mg./kg. (LD Rats 0.126'mg./kg. (2 LD Rabbits:

Intravenous 0.340 mg./kg. (20 LD Subcutaneous 0.900 mgJkg. (20 LD Dogs and cats:

Intravenous 0.440 mgjlcg. (20 LD Subcutaneous 0.900 rug/kg. (20 LD To minimize absorption eilects both the GB and TAM- 4 were ordinarily given intravenously. However, in the therapeutic tests on rabbits, dogs and cats, the GB was administered subcutaneously, since death from 20 LD intravenous dose of GB occurs so quickly that it is virtually impossible to give timely administration of the antidote.

Atropine, when administered, was included in the following amounts.

10 R'ats 4 Rabbits 2 Dogs and cats 0.5

The prophylactic doses were given within two minutes prior to the injection of the GB, the therapeutic doses so soon as poisoning symptoms were visible.

Table 3 shows the results.

Table 3 A. PROPHYLATIC 2-PAM TMB-4 Survival ratio Survival ratio Animals Animals 25 Dose Dose mgJkg. With- With mgJkg. With- With out atroout atroatropine atropine pine pine Mice 40 12 0/10 Rats 25 0/6 Rabbi-.- 5 6 2/5 Oats 40 2O 5/5 Dogs 4U 20 5/5 The recovery periods, i.e., time for disappearance of symptoms of poisoning, among survivors in the above tests, with atropine, were as follows.

4.5 Table 4 2-PAM TMB-4 Aminals Prophylactic Therapeutic Prophylactic Therapeutic Rats min 15 min Rabbits. 3 hr 30 min 2 hr.

Cats 5 hr. 5 hr 5 hr 24 hr.

Dogs- 24 hr. 24 hr. 1%; hr 3 hr.

The compounds of Formula I in which R contains from 7 to 10 carbon atoms are less effective than those of our preferred group. For these compounds the reactivation rate constant and the survival ratio for rats (measured as given above) were as follows, X- being bromide.

Table 5 R Rate constant Survival rate (BG) 2x10 0 4 i852 1. 2x10 olr (CHM (0119 0 1. 4x10 0/6 While these compounds were ineffective in vivo against GB, they were, together with 2-PAM, very ellective against certain other anticholinesterases, particularly that designated as VX by the US. Army Chemical Corps. All these compounds caused survival of all animals (survival rates of 4/ 4 and 6/ 6) when administered therapeutically to rats challenged by 2 LD doses of VX.

When X- was chloride the compound had the following properties. (In this and all following tables the survival ratios are those for rats challenged by 2 LD doses or GB or (VX) and the oxime was employed therapeutically.)

Compounds of Formula 11 showed reactivation rates very close to those of our preferred group. Thus when X- in Formula III is bromide the reactivation rate constant was 8X10 as compared to the value for the R: (CH member of our preferred group of 6x10 For the unsaturated member (Formula 111) the survival ratio for rats challenged by GB was only 1/4 as compared to 6/6 for the saturated analogue (Formula I). Both gave complete survival (ratios of 4/4 and 6/ 6) for animals challenged by VX, however.

Compounds of Formula IVa showed anomalous properties.

They gave reactivation rates which were low, but survival ratios which were high as compared to Z-PAM, as shown by Table 6, X- being bromide.

Table 6 R Reactivation Survival rate constant ratio (GB) (CH2)a- 3. 5X10 4/4 (CHz)5 4. 2X10 3/4 The compounds of group IVb, which are closely related to those of IVa, were somewhat less effective. When X- was bromide the compound had the following properties: Reactivation rate constant 2x10 survival ratio (GB) 2/4.

The compounds of group V were another group in which the results of therapeuatic treatment against GB were better as compared to Z-PAM then the reactivation rate constants would suggest, as shown by the following table, X- being bromide.

The compounds of Formulas VI, VII and VIII, while being of diflerent structure are alike in exhibiting reactivation rate constants which are very low as compared to 2-PAM but giving high survival ratios as shown by Table 8, X- being bromide in each case.

Table 8 Formula N0. Reactivation Survival rate ratio (GB) VT 69 4/4 VII Negligible 4/4 VTTI' 67 4/4 PREPARATION 011" COMPOUNDS 4-pyridinecarboxaldehyde oxime was prepared by warming on a steam bath a neutralized aqueous solution of 4-pyridinecarboxaldehyde and hydroxylamine hydrochloride. The oxime had a melting point of 130-1305 C. The 2- and 3-oximes were produced by similar methods.

The quaternization to produce dioximes was carried out by reacting the proper oxime with a 1, n dihaloalkane, (CH X employing a 3:1 molar ratio of oxime to halide. The unsymmetrical quaternary monoximes were obtained by reacting the pyridine oxime with the appropriate omega-halopropyl quaternary salt in a 15:1 molar ratio. Two procedures were utilized.

Procedure A.-A mixture of the pyridine oxime and halide was dissolved in sufiicient ethanol and refluxed for the period of time specified in Table 9.

Procedure B.-A mixture of the oxime and halide was dissolved in about ml. of ethanol and heated in a 200 ml. capped pressure bottle (carbonated beverage type) for the length of time specified. The reaction mixtures were cooled to room temperature and the product removed by filtration. In several instances it was necessary to add absolute ether to effect complete precipitation. The products were recrystalized from ether. This procedure was usually employed because of its simplicity.

Table 9 gives the procedure, yields and melting points for representative compounds.

Table 9 H H acoocsqooeooooooe s s s z-s s Further details regarding the preparation and properties of certain of our compounds are given in the following publications: by us and our associates:

Pyridine Aldoximes, by Edward J. Poziomek, Brennie E. Hackley, Jr. and George M. Steinberg, Journal of Organic Chemistry, vol 23, pp. 714-717 (May 1958); and Chemotherapeutic Elfectiveness of Trimethylene Bis (4-Formyl Pyridinium Bromide) Dioxime in Anticholinesterase Poisoning, by Edmund Bay, S. Kropp and L. F. Yates, Proceedings of the Society for Experimental Biology and Medicine, vol. 98, pages 107-109 (May 1958). These articles are to be considered incorporated by reference in this specification.

While we have shown a number of specific examples of compounds and their use, it will be obvious that various changes can be made without departing from our invention, which is defined by the following claim.

We claim:

A 1,1-trimethylene bis-(S-amidoxime pyridinium) halide.

No references cited: 

